ObJective To study the effects of flurbiprofen axetil preemptive analgesia on T cell immune functions in postoperative patients with upper abdominal.Methods Seventy-six patients with upper abdominal operation were divided into observation group (40 cases) and control group (36 cases) by random digits table,observation group and control group were given intravenous injection of 100 mg flurbiprofen axetil or 10 ml 0.9％ sodium chloride 10 min before skin incision respectively.Peripheral venous blood samples were collected for the determination of CD4+,CD8+,natural killer cell (NK cell) and CD4+/CD8+ by flow cytometry 1 day preoperative,2 and 48 h postoperative.Results In two groups,the CD8+ and NK cell of 2 h postoperative were lower significantly than those of preoperative (observation group:0.1850 ±0.0550 vs.0.2430 ±0.0856,0.1197 ±0.0673 vs.0.1598 ±0.0775；control group:0.1219 ±0.0571 vs.0.2385 ±0.0847,0.0778 ± 0.0311 vs.0.1621 ± 0.0806,P ＜ 0.05),however,the observation group was significantly higher than the control group (P ＜ 0.05).There was no significant difference in CD4+,CD8+ and NK cell of 48 h postoperative between two groups.Conclusion There are disorders on the T cell immune functions in postoperative patients with upper abdominal,but flurbiprofen axetil preemptive analgesia can improve the T cell immune function.

Objective To investigate the effects of ?-interferon (IFN-? on the expression of costimulatory molecules B7-1 (CD80) and B7-2 (CD86) and Fas/FasL in human retina. Methods Nine human eyes were obtained from the eye-bank of Zhongshan Ophthalmic Center. Six eyes were used for making retinal wholemounts, and 12 retinal wholemounts from each eye were put into the 24-hole culture board which had 2ml DMEM/F12 culture medium in each hole. The wholemounts were divided into 3 groups whose concentration of IFN-? was 0, 200, and 1 000 U/ml respectively. After cultured in 37℃ culture box (95%O2,5%CO2) for 24 hours, the expressions of B7-1 (CD80), B7-2 (CD86), and Fas/FasL on these retinal wholemounts were detected by immunohistochemical method. The retinal wholemounts from 3 healthy people were detected by immunohistochemical method as the control. Results Expression of FasL but not B7-1, B7-2 or Fas was found in the control group, while the expression of B7-1, B7-2 and Fas and increased expression of FasL were found after cultured with IFN-?. Conclusion IFN-? may be involved in the occurrence of ocular immune response and induction of apoptosis via the stimulation of expression of costimulatory molecules and Fas/FasL, which plays an important role in the activation of T lymphocytes.

ObjectiveTo investigate the expression of costimulatory molecules, Fas/FasL, and major histocompatibility complex (MHC)-class Ⅱ antigens in normal ocular tissues.MethodsTwelve eyes were obtained from the eye bank of Zhongshan Ophthalmic Center within 16 to 24 hours postmortem. Six eyes were used for making the retinal wholemounts, and the tissues (iris and ciliary body, choroid, and retina) of the others were used for making the frozen sections. Immunohistochemical staining was carried out on these retinal wholemounts as well as on tissue sections to investigate the exprenion of B7-1 and B7-2 (costimulatory molecules), HLA-DR(MHC class Ⅱ), CD68 (macrophages), Fas/FasL.ResultsThe results of immunohistochemical staining showed the presence of B7-2, FasL, CD68 and HLA-DR in the iris and ciliary body. Expression of B7-1, FasL, CD68, and HLA-DR was found in the choroid while HLA-DR, CD68 and FasL were detectable in the retina.ConclusionExpression of costimulatory molecules, MHC-class Ⅱ molecules and molecules related to apoptosis is different in the iris, ciliary body, choroid, and retina, which may play an important role in the stability of the immunological microenvironment of these tissues.

Objective To study the distribution and drug resistance of pathogens in neonatal septicemia in order to provide clinical guidance for antibiotic usage.Methods This retrospective study analyzed blood culture and clinical data from 83 confirmed neonatal septicemia patients and the blood collection cultures were analyzed.Results A total of 84 strains were isolated from 83 cases of blood specimens,Gram positive bacteria,Gram negative bacteria and fungi were 38(45.2%),41(48.8%),5(6.0%),respectively.Gram positive bacteria was mainly coagulase negative staphylococcus and staphylococcus aureus,which were 13(15.5%) and 8(9.5%) respectively.Gram negative bacteria was mainly Escherichia coli and Klebsiella pneumonia,which were 25(29.8%) and 9(10.7%) respectively.Gram positive bacteria were found high resistance to penicillin G,amoxicillin clavulanate potassium,oxacillin and clindamycin,from 34.2% to 73.7%,but they were sensitive to vancomycin,teicoplanin and linezolid.Gram negative bacteria were found high resistance to ampicillin(82.9%),the constituent ratio of the extended spectrum βlactamases(ESBLs) was 34.1%,carbapenem resistant strains was not found.All fungi were sensitive to azoles.Conclusion Gram negative bacteria are the major pathogens in neonatal septicemia,with high infection rate of Escherichia coli and high constituent ratio of the ESBLs,and antimicrobial agents should be chosen according to blood culture and antimicrobial susceptibility results.

New-onset refractory status epilepticus is a rare and special clinical manifestation with high mortality. About half of the patients have no clear cause. At present, the pathogenesis is unclear, and the treatment plan is controversial. In recent years, it has been found that inflammatory and immune responses of the body may be involved in the pathogenic process, and it is called “inflammatory-immune mediated epileptic encephalopathy” based on the perspective of pathogenesis. There have also been many treatment attempts based on the inflammatory and immunological mechanisms, some of which have achieved satisfactory results. However, most of them are based on the review of small sample cases, and relevant guidelines are still lacking at present. In this paper, the definition, etiology, pathogenesis, clinical manifestations and treatment of persistent status of new-onset refractory status epilepticus are reviewed.

Objective To evaluate the different value of color Doppler ultrnsonography and Doppler vascular examinations in diagnosis for deep venous thrombosis(DVT)in the lower extremities.Methotis Imaging of color Doppler ultrasound scanning was employed as diagnostic criteria for DVT on 178 lower extremities of 146 suspected patients,as compared to the result by Doppler vascular examinations.Results Color Doppler ultrasonograph showed hish accuracy in diagnosis for DVT,as compared to that by Doppler vascular examination with 97.9 percent(142/145)positive for the femoral and popliteal veins and relatively lower positive diagnostic vallie for thrombosis in the inferior vena cava,iliac vein,anterior tibial vein,posterior tibial vein and calf veins.Conclusions Color Doppler ultrasonography is superior to Doppler vascular examination in determining DVT of the lower extremities and can be used as a main diagnostic method for it.Doppler vasculiar examination can be used as an initial screening method for DVT and deep venous angiography should not be used as a routine diagnostic measure for it.

Objective To evaluate the diagnostic role of color Doppler ultrasonogarphy(CDU),Doppler ultrasound(DU)in the diagnosis of lower extremity deep venous thrombosis(DVT). Methods In this study,84 patients(92 lower extremities)of lower extremity DVT were underwent CDU and DU and lower extremity deep venous angiography respectively. Results Total consistent rate,sensitivity,specificity,omission diagnostic rate,mistake diagnostic rate,Youden index,Odd product.positive predictive value,negative predictive value and Kappa of CDU in diagnosing lower extremity DVT was respectively 96.7%,95.7%,97.8%,4.3%,2.2%,0.935,990.0,97.8%.95.7%and 0.935(P=0.037).As Kappa of CDU(0.935)＞0.75 and its P(0.037)＜0.05.CDU can theoreticallv substitute for deep venous angiography;Above-mentioned indexes of DU were respectively 89.1%,87.2%,91.1%,12.8%,8.9%,0.783,70.0,91.1%,87.2%and 0.783(P=0.065). Conclusion CDU iS becoming preferred and more reliable noninvasive method in diagnosing lower extremitv DVT.

BACKGROUND: In the field of intervertebral disc tissue engineering, seed cells primarily consist of nucleus pulposus cells (NPCs) and mesenchymal stem cells (MSCs). NPCs have been known to have several shortcomings: limited source, inconvenient collection, poor proliferative capacity, and difficult in vitro culture.OBJECTIVE: To investigate the proliferation of MSCs after co-culture with NPCs in alginate beads-simulated three-dimensional environment.DESIGN, TIME AND SETTING: A cell observation was performed at the Laboratory of Orthopedics, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology between December 2007 and October 2008.MATERIALS: Six healthy New Zealand rabbits of 4 months old were provided by the Laboratory Animal Center, Tongji Medical College of Huazhong University of Science and Technology and were used for the present study.METHODS: Rabbit MSCs were isolated and purified using density gradient centrifugation and adherence methods. Rabbit NPCs were isolated and purified using collagenase Ⅱ digestion and adherence methods. Following liposome-mediated green fluorescent protein tranfection and G418 screening, MSCs of passage 3 were cultured either alone (control group) or with NPCs at a ratio of 1:1 (co-culture group) in alginate beads. After 14 days of culture, alginate beads were dissolved and MSCs were collected by flow cytometry sorting.MAIN OUTCOME MEASURES: The expression of collagen Ⅱ and aggrecan in MSCs was detected by RT-PCR and Western blot techniques.RESULTS: mRNA and protein expression of collagen Ⅱ and aggrecan was observed in the co-culture group, but not in the control group, after 14 days of culture.CONCLUSION: MSC differentiation towards nucleus pulposus-like cells can be induced by co-culture with NPCs in a three-dimensional environment.

OBJECTIVE To investigate the effect of furanodiene(FDE),a diterpene derived from the medicinal plant Zedoary,on apoptosis of human gastric cancer MGC-803 cells induced in vitro. METHODS MGC-803 cells were treated with FDE 46.29~740.74μmol·L-1 for 24,48 and 72 h,and the cell viability was detected with MTT assay. Cell morphology was observed by light microscopy and Hoechst33342 staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. Rh123 staining and fluorescence probe DCFH-DA were employed to detect the changes in mitochondrial membrane potential (MMP) and reactive oxygen species(ROS). RESULTS MTT Results showed that FDE 46.29-740.74μmol · L-1 exhibited significantly higher cytotoxicity to gastric cancer MGC-803 cells. IC50 for MGC-803 of 24,48 and 72 h treatment was 347.91,257.41 and 101.01μmol·L-1,respectively. Treatment with FDE 92.58-370.32μmol·L-1 for 24 h also caused significant morphological changes in MGC-803 cells. AnnexinⅤ-FITC/PI double staining showed that the apoptotic rate increased after FDE 92.58-370.32μmol·L-1 treatment for 24 h(P<0.05). FDE enabled MGC-803 cell cycle arrest in S phase. DCFH-DA staining showed that FDE resulted in an increase in intracellular ROS levels(P<0.05) when PDE concentration was 370.37μmol·L-1(P<0.05). MMP decreased after FDE treatment when PDE concen?tration was 370.37μmol·L-1(P<0.05). CONCLUSION FDE Possesses potent tumor selected toxicity and can induce apoptosis of MGC-803 cells through cell cycle arresting,which is related to inhibition of DNA biosynthesis.

Objective To study the effects of small dosage insulin on intestinal inflammatory responses to endotoxin rats. Method Thirty two male SD rats were randomly divided into 4 groups (n = 8): control group, endotoxin (LPS,6 mg/kg i.p.)group, regular insulin(RI,0.5 IU/kg hypodermic) group and LPS(6 mg/kg i.p) + RI (0.5 IU/kg hypodermic)group. Six hours after LPS or saline injection,all rats were laparotomized to observe the congestion in intestinal mucosa with naked-eyes and photography.Then a segment of intestine was stained with HE to observe the pathological changes. The expressions of IL-6 and TNF-α were detected by RT-PCR.The systemic inflammatory response,blood sugar and food taken in rats were observed simultaneously. Software SPSS 13.0 was used to perform ANOVA and Chi-square test for statistical analysis. Results Compared with LPS group, the differences in hyperemia and inflammatory cell infiltration in intestinal tissue were not noticeable in LPS + RI group. The expression of IL-6 and TNF-α were significantly attenuated in RI + LPS group (P < 0.01). All rats in LPS group manifested systemic inflammatory response syndrome (SIRS) four or five hours after LPS treatment, while there was none in LPS + RI group. Rats in LPS group took less food than rats of other groups while the blood sugar had litter difference in all groups (P > 0.05). Conclusions Small dosage of insulin could reduce intestinal inflammation caused by endotoxemia. Early administration of insulin ould prevent the presence of SIRS while it has no obvious influence on blood sugar.